Increased Candida Demonstrated in Subgingival Plaques of Aids Patients by Microwave-Accelerated Silver Staining

1990 ◽  
Vol 189 ◽  
Author(s):  
Jacob S. Hanker ◽  
Mark J. Kutcher ◽  
E. Jefferson Burkes ◽  
George W. Greco ◽  
Roy L. Hopfer ◽  
...  
Keyword(s):  
Periodontal Disease ◽  
Oral Candidiasis ◽  
Periodic Acid ◽  
Rapid Test ◽  
Acid Oxidation ◽  
Subgingival Plaque ◽  
Gram Negative Bacteria ◽  
Aids Patients ◽  
Gram Negative ◽  
Large Numbers

ABSTRACTAcute periodontal lesions generally respond to conventional therapy. In AIDS patients, however, further deterioration is the usual consequence. Candida albicans(CA) lesions of the oral mucosa have been observed in 88% of AIDS patients; 59% of male homosexual and IV drug abuser oral candidiasis patients subsequently developed AIDS. Indeed HIV may be hard to find in AIDS patients in whom CA lesions are very prominent.Rapid reproducible silver stains have been developed in our laboratories to detect Gram-negative bacteria, fungi and protozoa. They demonstrate these microorganisms by the microwave-accelerated metallization of the aldehydes produced in their surface carbohydrates or lipopolysaccharides after periodic acid oxidation (Giammara and Hanker, US Patent No. 4,690,901, 1987; Sigma Diagnostics Kit No. HT-100). We thought that these stains might be useful to demonstrate by microscopy the large numbers of CA which culture studies had suggested were present in subgingival plaque at atypical periodontal disease sites of AIDS patients. Direct staining of plaque smears and their evaluation under the light microscope could be done in much less time than that required for culture. Any equivocal microorganisms could be positively identified as CA by immediate examination of the slide or coverslip by scanning electron microscopy. Although our PATS reaction was more valuable for identifying Gram-negative bacteria especially spirochetes and for studying substructure of CA and their epiphytic interactions with bacteria, the SIGMA DIAGNOSTICS Silver Stain HT-100 was much easier to use for rating CA concentration in disease sites. This test was applied to subgingival plaque smears from 28 periodontal disease sites of 22 HIV-seronegative patients and 45 sites of 12 HIV-seropositive patients. After staining, the smears were examined and scored (Davenport Index). None of the 12 HIVseropositive patients but 15 of the 22 HIV-seronegative patients had a score of zero at all sites. The score ranged from 0 (no CA) to 3 (large numbers of CA in all fields). In the seropositive patients the median score was 2 while in the seronegative group it was 0.The much higher scores obtained in specimens from periodontal disease sites of the AIDS patients suggest that this simple rapid test might be valuable for screening individuals and indicating those requiring further testing or monitoring for AIDS.

Neutrophil pseudoplatelets in subgingival plaque and crevicular fluid samples from periodontal diseased sites

1989 ◽  
Vol 47 ◽  
pp. 862-863 ◽  
Author(s):  
J Hanker ◽  
E.J. Burkes ◽  
G. Greco ◽  
R. Scruggs ◽  
B. Giammara
Keyword(s):  
Electron Microscopy ◽  
Bone Marrow ◽  
Peripheral Blood ◽  
Gingival Crevicular Fluid ◽  
Light And Electron Microscopy ◽  
Subgingival Plaque ◽  
Staining Reaction ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Crevicular Fluid

The mature neutrophil with a segmented nucleus (usually having 3 or 4 lobes) is generally considered to be the end-stage cell of the neutrophil series. It is usually found as such in the bone marrow and peripheral blood where it normally is the most abundant leukocyte. Neutrophils, however, must frequently leave the peripheral blood and migrate into areas of infection to combat microorganisms. It is in such areas that neutrophils were first observed to fragment to form platelet-size particles some of which have a nuclear lobe. These neutrophil pseudoplatelets (NPP) can readily be distinguished from true platelets because they stain for neutrophil myeloperoxidase. True platelets are not positive in this staining reaction because their peroxidase Is inhibited by glutaraldehyde. Neutrophil pseudoplatelets, as well as neutrophils budding to form NPP, could frequently be observed in peripheral blood or bone marrow samples of leukemia patients. They are much more prominent, however, in smears of inflammatory exudates that contain gram-negative bacteria and in gingival crevicular fluid samples from periodontal disease sites. In some of these samples macrophages ingesting, or which contained, pseudoplatelets could be observed. The myeloperoxidase in the ingested pseudoplatelets was frequently active. Despite these earlier observations we did not expect to find many NPP in subgingival plaque smears from diseased sites. They were first seen by light microscopy (Figs. 1, 3-5) in smears on coverslips stained with the PATS reaction, a variation of the PAS reaction which deposits silver for light and electron microscopy. After drying replicate PATS-stained coverslips with hexamethyldisilazane, they were sputter coated with gold and then examined by the SEI and BEI modes of scanning electron microscopy (Fig. 2). Unstained replicate coverslips were fixed, and stained for the demonstration of myeloperoxidase in budding neutrophils and NPP. Neutrophils, activated macrophages and spirochetes as well as other gram-negative bacteria were also prominent in the PATS stained samples. In replicate subgingival plaque smears stained with our procedure for granulocyte peroxidases only neutrophils, budding neutrophils or NPP were readily observed (Fig. 6).

scholarly journals Could Periodontal Disease through Periopathogen Fusobacterium nucleatum Be an Aggravating Factor for Gastric Cancer?

2020 ◽  
Vol 9 (12) ◽  
pp. 3885
Author(s):  
Petra Șurlin ◽  
Flavia Mirela Nicolae ◽  
Valeriu Marin Șurlin ◽  
Ștefan Pătrașcu ◽  
Bogdan Silviu Ungureanu ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Periodontal Disease ◽  
Anaerobic Bacterium ◽  
Fusobacterium Nucleatum ◽  
Negative Influence ◽  
Chronic Inflammatory Disease ◽  
Periodontal Pathogen ◽  
Future Research ◽  
Gram Negative Bacteria ◽  
Gram Negative

Periodontal disease affects the supporting tissues of the teeth, being a chronic inflammatory disease caused by specific microorganisms from subgingival biofilm. Fusobacterium nucleatum is a Gram-negative anaerobic bacterium that acts as a periodontal pathogen, being an important factor in linking Gram-positive and Gram-negative bacteria in the periodontal biofilm, but its involvement in systemic diseases has also been found. Several studies regarding the implication of Fusobacterium nucleatum in gastro-enterological cancers have been conducted. The present review aims to update and systematize the latest information about Fusobacterium nucleatum in order to evaluate the possibility of an association between periodontal disease and the evolution of gastroenterological cancers through the action of Fusobacterium nucleatum, highlighting gastric cancer. This would motivate future research on the negative influence of periodontal pathology on the evolution of gastric cancer in patients suffering from both pathologies.

scholarly journals Evaluation of an overnight non-culture test for detection of viable Gram-negative bacteria in endoscope channels

2019 ◽  
Vol 07 (02) ◽  
pp. E268-E273 ◽  
Author(s):  
Harminder Singh ◽  
Donald Duerksen ◽  
Gale Schultz ◽  
Carol Reidy ◽  
Pat DeGagne ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Rapid Test ◽  
Gram Negative Bacteria ◽  
Clinical Workflow ◽  
Gram Negative ◽  
E Coli ◽  
Test Kit ◽  
Viable Bacteria ◽  
Low Levels ◽  
High Level

Abstract Background and study aims Prevention of infection transmission from contaminated endoscopes would benefit from a rapid test that could detect low levels of viable bacteria after high level disinfection. The aim of this study was to evaluate the rapid NOW! (RN) test’s ability to detect endoscope contamination. Materials and methods The RN test kit and the accompanying fluorometer were evaluated. The manufacturer states that a fluorometer signal > 300 units is indicative of viable Gram-negative bacteria. Suspension testing of varying concentrations of Escherichia coli, Pseudomonas aeruginosa and Enterococcus faecalis were used to determine the RN test limit of detection. Simulated-use testing was done using a duodenoscope inoculated with 10 % blood containing approximately 35 CFU E. coli per channel. Samples were extracted from the duodenoscope instrument channel and tested using the manufacturer’s instructions. Results The RN test could consistently detect 10 CFU of E. coli and P. aeruginosa (fluorescent signal of 9,000 to 11,000 units) but not E. faecalis. Sensitivity and specificity for Gram-negative bacteria were 93 % and 90 %, respectively, using all of the suspensions in the study. Extraction of E. coli from an inoculated duodenoscope instrument channel repeatedly provided a positive signal (i. e. > 2,000 units). Conclusions The RN test can reliably detect low levels of Gram-negative bacteria in suspension as well as from samples extracted from endoscope channels. These preliminary findings are encouraging but further assessment of extraction efficacy, impact of organic residuals and clinical workflow are still needed.

scholarly journals pET expression vector customized for efficient seamless cloning

BioTechniques ◽  
2020 ◽  
Vol 69 (5) ◽  
pp. 384-387
Author(s):  
Dragana Dobrijevic ◽  
Lily A Nematollahi ◽  
Helen C Hailes ◽  
John M Ward
Keyword(s):  
Expression Vector ◽  
Gene Replacement ◽  
Expression Vectors ◽  
Gram Negative Bacteria ◽  
Golden Gate ◽  
Gram Negative ◽  
Large Numbers ◽  
Parallel Cloning ◽  
Natural Enzyme

Here we present a modification of the widely used pET29 expression vector for use in rapid and straightforward parallel cloning via a gene replacement and Golden Gate strategy. The modification can be applied to other expression vectors for Gram-negative bacteria. We have used the modified vectors to clone large numbers of bacterial natural enzyme variants from genomic and metagenomic sources for applications in biocatalysis.

scholarly journals Photoactive Porphyrin Derivative with Broad-Spectrum Activity against Oral Pathogens In Vitro

2000 ◽  
Vol 44 (12) ◽  
pp. 3364-3367 ◽  
Author(s):  
C. R. Rovaldi ◽  
A. Pievsky ◽  
N. A. Sole ◽  
P. M. Friden ◽  
D. M. Rothstein ◽  
...  
Keyword(s):  
Periodontal Disease ◽  
Pathogenic Bacteria ◽  
Vitro Activity ◽  
Chlorin E6 ◽  
Periodontal Pocket ◽  
Gram Negative Bacteria ◽  
In Vitro Activity ◽  
Gram Negative ◽  
Porphyrin Derivative

ABSTRACT Photodynamic therapy (PDT) has historically been used as a means to treat cancerous tumors but has recently been used to kill bacterial cells through the use of targeted photosensitizers. PDT is a potential adjunct to scaling and root planing in the treatment of periodontal disease. However, the effectiveness of porphyrin derivatives against microorganisms has been limited because some gram-negative bacteria are refractory to photodynamic treatment with these agents. We have designed a porphyrin derivative conjugated to a pentalysine moeity that endows the molecule with activity against gram-positive and gram-negative bacteria. Whereas the porphyrin, chlorin e6, showed in vitro activity against a limited spectrum of bacteria, chlorin e6 conjugated to pentalysine showed in vitro activity against all oral microorganisms tested, including Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatumsubsp. polymorphum, Actinomyces viscosus, and the streptococci. Potent antimicrobial activity (≥5-log-unit reduction in the numbers of CFU per milliliter) was retained in the presence of up to 25% whole sheep blood. The use of potent, selective agents such as this chlorin e6–pentalysine conjugate to more effectively reduce the pathogenic bacteria in the periodontal pocket may be a significant tool for the treatment of periodontal disease.

scholarly journals Exogenous polyunsaturated fatty acids (PUFAs) promote changes in growth, phospholipid composition, membrane permeability and virulence phenotypes in Escherichia coli

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Joshua L. Herndon ◽  
Rachel E. Peters ◽  
Rachel N. Hofer ◽  
Timothy B. Simmons ◽  
Steven J. Symes ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fatty Acids ◽  
Fatty Acid ◽  
Membrane Permeability ◽  
Phospholipid Composition ◽  
Acid Oxidation ◽  
Gram Negative Bacteria ◽  
Membrane Phospholipids ◽  
Gram Negative ◽  
E Coli

Abstract Background The utilization of exogenous fatty acids by Gram-negative bacteria has been linked to many cellular processes, including fatty acid oxidation for metabolic gain, assimilation into membrane phospholipids, and control of phenotypes associated with virulence. The expanded fatty acid handling capabilities have been demonstrated in several bacteria of medical importance; however, a survey of the polyunsaturated fatty acid responses in the model organism Escherichia coli has not been performed. The current study examined the impacts of exogenous fatty acids on E. coli. Results All PUFAs elicited higher overall growth, with several fatty acids supporting growth as sole carbon sources. Most PUFAs were incorporated into membrane phospholipids as determined by Ultra performance liquid chromatography-mass spectrometry, whereas membrane permeability was variably affected as measured by two separate dye uptake assays. Biofilm formation, swimming motility and antimicrobial peptide resistance were altered in the presence of PUFAs, with arachidonic and docosahexaenoic acids eliciting strong alteration to these phenotypes. Conclusions The findings herein add E. coli to the growing list of Gram-negative bacteria with broader capabilities for utilizing and responding to exogenous fatty acids. Understanding bacterial responses to PUFAs may lead to microbial behavioral control regimens for disease prevention.

scholarly journals Microbiological Aspect in Dental Implants and Natural Teeth with the Impact of Aloe vera Gel

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Batool Hassan AL-Ghurabi ◽  
Ahlam T Mohammed ◽  
Nadia Aftan AL-Rawi
Keyword(s):  
Dental Implants ◽  
Aloe Vera ◽  
Subgingival Plaque ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Aloe Vera Gel ◽  
Natural Teeth ◽  
Microbiological Aspect ◽  
The Impact

The health of peri-implant tissues is one of the most important aspects for the long-term success of dental implants. Thus the control of bacterial plaque which is the main etiological factor in peri-implant diseases by using an effective preventive regimen and a therapeutic agent is of vital importance in prevention of any irreversible pathological conditions of peri‑implant tissues. The purposeof this study wasto investigate and compare microbiological aspect of subgingival plaque at implants and contralateral natural teeth in the same patients with the impact of Aloe vera gel. 20 females aged 25- 45 year who exhibited the presence of one clinically stable implant carrying a full ceramic crown and one contralateral natural tooth were selected to be enrolled in a split-mouth study. All participants were apparently healthy with no history of any systematic diseases and they have been treated with a two-piece implant system which should be in function for at least 6 months. Samples of subgingival plaque were collected from implants and contralateral teeth for microbial analyses (Gram positive bacteria, Gram negative bacteria and Candida species) at base line and 2 weeks after the application of Aloe vera gel. Microbial analyses revealed no statistically significant difference (p> 0.05) in the counts of microorganisms between implants and teeth, although they were lower in implant than that in teeth. Finding also revealed a significant reduction (p< 0.01) in the count of Gram-positive bacteria and Candida speciesafter application of Aloe vera for both implants and teeth. On the other hand, the count of Gram-negative bacteria seems not to be affected by Aloe vera gel. No differences were found in microbiological analysis of subgingival plaque from dental implants and teeth in the same patient. Moreover, Aloe vera is a promising herb with antimicrobial activity that could be used as a beneficial agent in treatment and prevention of peri-implant diseases.

Elevated Candida albicans scores obtained by microscopic examination of subgingival plaque from atypical periodontal disease sites in immunocompromised states (especially AIDS)

1990 ◽  
Vol 48 (3) ◽  
pp. 616-617 ◽  
Author(s):  
J. Hanker ◽  
W.P. Webster ◽  
E.J. Burkes ◽  
G. Greco ◽  
D. Hamamoto ◽  
...  
Keyword(s):  
Radiation Therapy ◽  
Candida Albicans ◽  
Periodontal Disease ◽  
Oral Mucosa ◽  
Microscopic Examination ◽  
Conventional Therapy ◽  
Relative Number ◽  
Subgingival Plaque ◽  
Aids Patients ◽  
Almost All

Gingivitis and periodontitis have immunologic aspects that can influence or be influenced by the pathogenic microbiota present. In immunocompromised states, especially AIDS, an abnormal gingivitis (HIV-G) and periodontitis (HIV-P) are known. Although acute periodontal lesions generally respond to conventional therapy, in AIDS patients the atypical lesions may also be present which deteriorate further. Candida albicans(CA) lesions of the oral mucosa (not due to diabetes, cancer chemotherapy, radiation therapy or antibiotics) were present in almost all AIDS patients and were helpful in defining this disease. In 1988 it was shown that CA could be cultured from a greater percentage of the atypical than the conventional gingivitis or periodontitis sites in AIDS patients but this required many hours and could not show the relative number of CA present in the different sites.

Demonstration of activated macrophages in subgingival plaque and crevicular fluid samples from periodontal diseased sites

1989 ◽  
Vol 47 ◽  
pp. 1084-1085 ◽  
Author(s):  
J. Hanker ◽  
E.J. Burkes ◽  
G. Greco ◽  
R. Scruggs ◽  
E. Anderson ◽  
...  
Keyword(s):  
Periodic Acid ◽  
Nonspecific Esterase ◽  
Subgingival Plaque ◽  
Activated Macrophages ◽  
Periodic Acid Schiff ◽  
Intense Staining ◽  
Gram Negative ◽  
Periodontal Tissues ◽  
Crevicular Fluid

Macrophages are present in gingival tissues and they increase in gingivitis and periodontitis. In vitro studies have shown that gram-negative bacterial endotoxin increases the bone resorption activity of macrophages. The activation of macrophages in vitro by substances such as lymphokines, complement, group A streptococcal wall substances, histamine and immune complexes which are present in subgingival plaque or periodontal tissues has also been shown. Intense staining of activated macrophages for acid phosphatase and nonspecific esterase has been observed in tissue sections of inflamed gingiva (cf. 4,9). Although macrophages have also been observed in crevicular fluid and subgingival plaques from normal sites, their numbers, enzyme activities or activation have not previously been studied in crevicular fluid and subgingival plaques from diseased sites. All of the following studies were performed on samples obtained by noninvasive procedures and smeared on coverslips. After cytochemical procedures which produced visible and electron opaque stains on replicate coverslips, correlative light and scanning electron microscopy by SEI and BEI modes permitted identification of cell types and some bacterial pathogens. The PATS reaction, a variation of the periodic acid-Schiff(PAS) reaction that employs stepwise condensation with thiocarbohydrazide and silver methenamine to achieve staining and electron opacity, has been found useful by us for positively staining gram-negative bacteria including spirochetes in crevicular fluid and subgingival plaques from periodontal patients. Neutrophils were intensely stained due to their glycogen content. Larger glycogen positive cells, frequently highly vacuolated, were also present; they appeared to be activated macrophages (Figs. 1-3).

Sign in / Sign up

Export Citation Format

Share Document